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Evaluation of agar gel double diffusion test for detection of nuclear polyhedrosis virus of silkworm, Bombyx mori L.

Published online by Cambridge University Press:  19 September 2011

J. Bhattacharya ,
N. Krishnan ,
A. K. Chandra ,
S. K. Sen  and
B. Saratchandra
J. Bhattacharya
Affiliation:
Central Sericultural Research and Training Institute, Berhampore, West Bengal, India–742 101
N. Krishnan
Affiliation:
Central Sericultural Research and Training Institute, Berhampore, West Bengal, India–742 101
A. K. Chandra
Affiliation:
Central Sericultural Research and Training Institute, Berhampore, West Bengal, India–742 101
S. K. Sen
Affiliation:
Central Sericultural Research and Training Institute, Berhampore, West Bengal, India–742 101
B. Saratchandra
Affiliation:
Central Sericultural Research and Training Institute, Berhampore, West Bengal, India–742 101
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Abstract

The agar gel double diffusion test was evaluated for detection of nuclear polyhedrosis virus infecting the silkworm, Bombyx mori L. (BmNPV). The test was successful using alkali-dissolved BmNPV as antigen. The antiserum raised against dissolved BmNPV gave positive results with a wider range of antigenic concentrations compared to antiserum raised against undissolved BmNPV. Gel plates prepared using phosphate buffered-saline as dissolving medium for agarose gave sharper and clearer precipitation bands compared to those made with sodium carbonate-saline as diluent. Sharp bands were obtained when the experiment was carried out at 25°C, whereas at higher temperature (35°C), the bands were diffused. The test was able to detect antigen concentration as low as 2 × 106 OBs/ml (18.9 μg/ml of antigen protein).

Résumé

Le test de double diffusion en gel d'agar a été évalué pour la détection du virus à polyhédrose nucléaire infectant le ver à soie, Bombyx mori L. (BmNPV). Le test a réussi en utilisant le BmNPV en solution basique comme antigène. L'antisérum produit centre le BmNPV dissous a donné des résultats positifs avec une plus large gamme de concentrations antigéniques par rapport à l'antisérum produit par le BmNPV non dissous. Les plaques de gel préparées en utilisant une solution saline en tempon phosphate comme milieu dissolvent pour l'agarose a donné des bandes de précipitation plus nettes et plus claires par rapport à celles produites avec le sel de phosphate de sodium comme diluant. De bandes nettes étaient obtenues quand l'expérience se déroulait à 25°C, alors qu'à plus haute température (35°C), les bandes étaient diffuses. Le test pouvait détecter l'antigène jusqu'à une concentration aussi basse que 2 × 106 OBs/ml (18.9 μg/ml de protéine antigénique).

Keywords

silkwormBombyx mori nuclear polyhedrosis virus (BmNPV) detectionagar gel double diffusion testBombyx moridetection du virus à polyhédrose nucléaire (BmNPV)test de double diffusion en gel d'agar
Type
Research Articles
Information
International Journal of Tropical Insect Science , Volume 16 , Issue 3-4 , December 1995 , pp. 351 - 356
Copyright
Copyright © ICIPE 1995

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References

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